三氧化二砷抑制小鼠乳腺癌细胞MA-891生长作用及其对端粒酶活性的影响  

Inhibitory mechanism on growth of MA-891 cells by arsenic trioxide

作　　者：石莹[1] 胡博[2] 郭俣[1] 王慧[3] 夏俊[1] 

机构地区：[1]蚌埠医学院生化与分子生物学教研室,安徽蚌埠233030 [2]苏州大学生物医学研究院肿瘤细胞与分子免疫实验室,江苏苏州215123 [3]蚌埠医学院临床检验诊断学实验中心,安徽蚌埠233030

出　　处：《中国中药杂志》2012年第5期637-642,共6页China Journal of Chinese Materia Medica

基　　金：安徽省教育厅自然科学研究项目(2003kJ256)

摘　　要：目的:探讨As2O3在诱导小鼠乳腺癌细胞MA-891凋亡过程中的作用,以及对端粒酶及其亚单位mTERT mRNA活性表达的影响,为进一步进行小鼠体内实验提供理论和实验依据。方法:以小鼠乳腺癌细胞株MA-891为研究对象,将不同浓度的As2O3与MA-891细胞共培养不同时间后,MTT法检测As2O3对MA-891细胞的生长抑制作用;流式细胞术检测As2O3对MA-891细胞凋亡的影响;TRAP-PAGE-银染法检测As2O3作用后MA-891细胞中端粒酶活性的改变;RT-PCR检测As2O3对MA-891细胞鼠端粒酶逆转录酶(mTERT)的影响。结果:As2O3能显著抑制MA-891细胞生长,24,48 h的半数生长抑制浓度(IC50)分别为9.68,5.50μmol.L-1。5,10,20μmol.L-1的As2O3诱导MA-891细胞24 h后,细胞凋亡率依次为30.21%,48.26%,57.43%。5,10,20μmol.L-1的As2O3作用MA-891细胞24,48 h后,端粒酶活性显著下降,抑制程度呈明显的时间和浓度依赖性;银染条带也显示出相同结果。RT-PCR产物显示mTERT mRNA表达显著下降并呈时间和浓度依赖关系。结论:As2O3能显著抑制MA-891细胞生长、诱导细胞凋亡,可能是通过抑制细胞端粒酶及其亚单位端粒酶逆转录酶的活性而发挥作用。Objective： To investigate the apoptosis in MA-891 cells induced by different concentrations of As2O3 and to study its influence on the activity of telomerase and telomerase mTERT-mRNA,which provide theoretical and experimental basis for breast carcinoma.Method： The MA-891 breast carcinoma cell lines were used as the object.Different concentration of As2O3 was cultivated with MA-891 cells,and the growth inhibition of MA-891 cells was analyzed by MTT.The rate of apoptosis in MA-891 cells was detected by flow cytometry.The telomerase activity was determined by TRAP-PAGE-SILVER staining and was analyzed by specific soft ware.The expression mTERT-mRNA was examined by RT-PCR assay in untreated and As2O3-treated cells.Result： As2O3 could inhibit the growth of MA-891 cells remarkably;the IC50 value of As2O3 for MA-891 was 9.68 μmol·L-1 and 5.50 μmol·L-1 respectively during 24,48 h.The percentage of the apoptosis in MA-891 cells was 30.21%,48.26%,57.43%,as the cells were treated with 5,10,20 μmol·L-1 As2O3 for 24 hours.Treated with As2O3 for 24 hand 48 h,the telomerase activity of MA-891 cells was inhibited remarkably,which showed obvious time and concentration dependence.The mTERT-mRNA expression of MA-891 cells were significantly inhibited when treated with As2O3 for 24 h and 48h.Conclusion： As2O3 could remarkably inhibit the growth of MA-891 cells and could promote the apoptosis of the cells.Treated with As2O3,the activities of telomerase and telomerase mTERT-mRNA were inhibited remarkably and were obvious dosage-effect correlation.

关 键 词：AS2O3 小鼠乳腺癌细胞MA-891 凋亡 端粒酶 端粒酶逆转录酶 

分 类 号：R737.9[医药卫生—肿瘤] R285.5[医药卫生—临床医学]